Compounds and methods for modulating target nuclear and sub-nuclear nucleic acid molecules in cells and animals

Abstract

The present invention provides compounds and methods for modulating target nucleic acids found in organelles or sub-organelles of cells. The invention includes, but is not limited to compounds and methods that modulate target nucleic acids in a sub-nuclear organelle, such as the nucleolus and/or a cajal body. In certain embodiments, the cell is in an animal.

1 . A method of reducing the amount or activity of a target snoRNA in a cell in an animal comprising administering to the animal a pharmaceutical composition comprising a single-stranded antisense compound comprising a modified oligonucleotide complementary to accessible nucleotides of the target snoRNA; and thereby reducing the amount or activity of the target snoRNA in the cell in the animal;
 wherein the modified oligonucleotide consists of 10 to 30 linked nucleosides,   wherein the modified oligonucleotide comprises:   a 5′-region consisting of 1 to 7 modified, linked nucleosides;   a 3′-region consisting of 1 to 7 modified, linked nucleosides; and   a central region consisting of 5 to 28 linked 2′-deoxyribonucleosides; and   wherein the animal is a mammal.   
     
     
         2 . The method of  claim 1 , wherein the modified oligonucleotide consists of 14 to 23 linked nucleosides, and the central region consists of 7 to 11 linked 2′-deoxyribonucleosides. 
     
     
         3 . The method of  claim 2 , wherein at least one modified nucleoside comprises a modified sugar moiety selected from among: 2′-MOE, 2′-OMe, 2′-F, and a BNA. 
     
     
         4 . The method of  claim 3 , wherein the BNA is selected from LNA, ENA, and cEt. 
     
     
         5 . The method of  claim 2 , wherein the modified oligonucleotide comprises one or more modified internucleoside linkages. 
     
     
         6 . The method of  claim 2 , wherein the antisense compound comprises a conjugate group. 
     
     
         7 . The method of  claim 2 , wherein the target snoRNA derives from a host RNA. 
     
     
         8 . The method of  claim 7 , wherein the amount and activity of the host RNA are essentially unchanged. 
     
     
         9 . The method of  claim 2 , wherein the target snoRNA is a C/D box snoRNA or an H/ACA box snoRNA. 
     
     
         10 . The method of  claim 9 , wherein the target snoRNA is a C/D box snoRNA. 
     
     
         11 . The method of  claim 9 , wherein the target snoRNA is an H/ACA box snoRNA. 
     
     
         12 . The method  claim 5 , wherein the one or more modified internucleoside linkages is a phosphorothioate internucleoside linkage. 
     
     
         13 . The method  claim 12 , wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate internucleoside linkage. 
     
     
         14 . The method  claim 2 , wherein the accessible nucleotides of the target snoRNA are accessible to dimethylsulphate in vitro. 
     
     
         15 . The method  claim 2 , wherein the modified oligonucleotide is at least 90% complementary to the accessible nucleotides of the target snoRNA. 
     
     
         16 . The method  claim 2 , wherein the modified oligonucleotide is 100% complementary to the accessible nucleotides of the target snoRNA. 
     
     
         17 . The method of  claim 2 , wherein each modified nucleoside comprises a modified sugar moiety independently selected from among: 2′-MOE, 2′-OMe, 2′-F, and a BNA. 
     
     
         18 . The method of  claim 2 , wherein the animal is a human. 
     
     
         19 . The method of  claim 2 , wherein the pharmaceutical composition is administered systemically. 
     
     
         20 . The method of  claim 2 , wherein the pharmaceutical composition is administered by subcutaneous injection.